| abstract |
Denatured proteins are renatured by (a) dissolving the denatured protein in a renaturing buffer to form a soln. with a predetermd. critical concn., (b) allowing time for a 'folding intermediate' to form, (c) adding more denatured protein in the amt. required to attain the critical concn., and (d) repeating steps (b) and (c) one or more times. The critical concn. is the concn. of denatured protein giving the max. yield of renatured protein after renaturation; its value is determd. in preliminary tests by plotting protein activity against initial concn. The 'folding intermediate' is a precursor of the renatured protein; the time required for its formation is determd. in preliminary tests, e.g. by plotting circular dichroism, fluorescence or viscosity against time. |